Total and Putative Surface Proteomics of Malaria Parasite Salivary Gland Sporozoites
Total and Putative Surface Proteomics of Malaria Parasite Salivary Gland Sporozoites
复制标题
DOI：
10.1074/mcp.m112.024505
复制
发表时间：
2013-05-01
影响因子：
7
通讯作者：
Kappe, Stefan H. I.
中科院分区：
文献类型：
--
作者：
Lindner, Scott E.;Swearingen, Kristian E.;Kappe, Stefan H. I.
哺乳动物的疟疾感染是由疟原虫唾液腺子孢子在按蚊媒介叮咬期间传播而引起的。子孢子穿过皮肤，最终到达肝脏，在那里它们感染肝细胞。阻断这一感染的初始阶段是一种很有希望的疟疾疫苗策略。因此，全面地阐明子孢子的蛋白质组成对于识别新的阻断感染的靶点将是非常有价值的。以前鉴定在蚊虫阶段表达的蛋白质的努力因将寄生虫与其媒介分离的技术困难而受阻；如果没有有效的纯化，所确定的绝大多数蛋白质都来自媒介。在这里，我们描述了来自两种疟原虫物种的高纯度唾液腺子孢子的蛋白质组学图谱：人感染恶性疟原虫和啮齿动物感染约氏疟原虫。改进的样品纯化和高质量精度质谱学的结合促进了迄今为止对该寄生虫红细胞前阶段最完整的蛋白质组覆盖。共鉴定了1991个恶性疟原虫子孢子蛋白和1876个约氏疟原虫子孢子蛋白，其中86%的蛋白具有较高的序列覆盖率。蛋白质组学数据证实了哺乳动物宿主感染子孢子的三个关键特征的存在：子孢子的运动和入侵装置(胶质小体)、子孢子信号通路和顶端分泌细胞器的含量。此外，活体子孢子上的蛋白质的化学标记和鉴定揭示了假定的子孢子表面暴露的蛋白质组以前未被表征的复杂性。综上所述，这些数据构成了迄今为止对唾液腺子孢子蛋白表达的最全面的分析，并揭示了潜在的潜在表面暴露蛋白，这些蛋白可能是抗体阻断感染的有价值的靶标。
Malaria infections of mammals are initiated by the transmission of Plasmodium salivary gland sporozoites during an Anopheles mosquito vector bite. Sporozoites make their way through the skin and eventually to the liver, where they infect hepatocytes. Blocking this initial stage of infection is a promising malaria vaccine strategy. Therefore, comprehensively elucidating the protein composition of sporozoites will be invaluable in identifying novel targets for blocking infection. Previous efforts to identify the proteins expressed in Plasmodium mosquito stages were hampered by the technical difficulty of separating the parasite from its vector; without effective purifications, the large majority of proteins identified were of vector origin. Here we describe the proteomic profiling of highly purified salivary gland sporozoites from two Plasmodium species: human-infective Plasmodium falciparum and rodent-infective Plasmodium yoelii. The combination of improved sample purification and high mass accuracy mass spectrometry has facilitated the most complete proteome coverage to date for a pre-erythrocytic stage of the parasite. A total of 1991 P. falciparum sporozoite proteins and 1876 P. yoelii sporozoite proteins were identified, with >86% identified with high sequence coverage. The proteomic data were used to confirm the presence of components of three features critical for sporozoite infection of the mammalian host: the sporozoite motility and invasion apparatus (glideosome), sporozoite signaling pathways, and the contents of the apical secretory organelles. Furthermore, chemical labeling and identification of proteins on live sporozoites revealed previously uncharacterized complexity of the putative sporozoite surface-exposed proteome. Taken together, the data constitute the most comprehensive analysis to date of the protein expression of salivary gland sporozoites and reveal novel potential surface-exposed proteins that might be valuable targets for antibody blockage of infection.